Derivation of Induced Pluripotent Stem Cells by Lentiviral Transduction

Nethercott, Hubert E.; Brick, David J.; Schwartz, Philip H.

doi:10.1007/978-1-61779-201-4_6

Cited by 18 publications

(15 citation statements)

References 7 publications

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“…In order to ensure uniformity among all the fibroblast lines, each of the fibroblast cell lines was derived by one group at CHOC Children's using documented standard operating procedures for media preparation (SBM, HDFM), skin biopsy processing, expansion, and cryopreservation. Additionally, all iPSCs were generated, expanded, and characterized according to published protocols by the same group of individuals [25, 32]. Multiple iPSC clones were characterized by pluripotent stem cell (PSC) morphology, uniformity of staining for classic PSC markers such as Tra‐1‐60, Nanog, Sox‐2, and Oct‐4 and normal karyotype (seven NSCs were also karyotyped and were shown to be normal similar to the iPSC from which they were derived; thus, this was not routinely done).…”

Section: Resultsmentioning

confidence: 99%

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The Autism Spectrum Disorders Stem Cell Resource at Children's Hospital of Orange County: Implications for Disease Modeling and Drug Discovery

Brick

Nethercott

Montesano

et al. 2014

Stem Cells Translational Medicine

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The autism spectrum disorders (ASDs) comprise a set of neurodevelopmental disorders that are, at best, poorly understood but are the fastest growing developmental disorders in the United States. Because animal models of polygenic disorders such as the ASDs are difficult to validate, the derivation of induced pluripotent stem cells (iPSCs) by somatic cell reprogramming offers an alternative strategy for identifying the cellular mechanisms contributing to ASDs and the development of new treatment options. Access to statistically relevant numbers of ASD patient cell lines, however, is still a limiting factor for the field. We describe a new resource with more than 200 cell lines (fibroblasts, iPSC clones, neural stem cells, glia) from unaffected volunteers and patients with a wide range of clinical ASD diagnoses, including fragile X syndrome. We have shown that both normal and ASD-specific iPSCs can be differentiated toward a neural stem cell phenotype and terminally differentiated into action-potential firing neurons and glia. The ability to evaluate and compare data from a number of different cell lines will facilitate greater insight into the cause or causes and biology of the ASDs and will be extremely useful for uncovering new therapeutic and diagnostic targets. Some drug treatments have already shown promise in reversing the neurobiological abnormalities in iPSC-based models of ASD-associated diseases. The ASD Stem Cell Resource at the Children's Hospital of Orange County will continue expanding its collection and make all lines available on request with the goal of advancing the use of ASD patient cells as disease models by the scientific community.

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Section: Resultsmentioning

confidence: 99%

“…Fibroblasts were reprogrammed to iPSCs using both traditional integrative (Lentivirus; 1IX) [32] and nonintegrative (Sendai virus; 2IX) methods [25].…”

Section: Methodsmentioning

confidence: 99%

The Autism Spectrum Disorders Stem Cell Resource at Children's Hospital of Orange County: Implications for Disease Modeling and Drug Discovery

Brick

Nethercott

Montesano

et al. 2014

Stem Cells Translational Medicine

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“…This demonstrates that the reprogramming procedure requires further optimization for the clinical applications of iPSCs and their derivatives. More recently, iPSCs have been derived with a lentivirus‐mediated approach and non‐integrated Sendai virus vectors with the hope to reduce these concerns . Additional approaches using RNAs, proteins, and chemical‐mediated reprogramming have been developed .…”

Section: Cell‐based Therapies For MImentioning

confidence: 99%

Stem Cell Therapy of Myocardial Infarction: A Promising Opportunity in Bioengineering

Jiang

Shamul

et al. 2020

Advanced Therapeutics

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Myocardial infarction (MI) is a life‐threatening disease resulting from the irreversible death of cardiomyocytes (CMs). Stem cell‐based therapies have been studied for MI treatment over the last two decades with promising outcomes. Here, the past work in this field is critically reviewed to elucidate the advantages and disadvantages of treating MI using pluripotent stem cells (PSCs) including both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), adult stem cells, and cardiac progenitor cells. Overall, PSCs (particularly iPSCs) have more advantages than the other cells for cardiac regeneration. However, the use of iPSCs is also facing critical challenges, which may be resolved by using biomaterials to engineer stem cells for reduced immunogenicity, improved immobilization/survival in the heart, and increased integration with the host cardiac tissue to eliminate arrhythmia. Biomaterials have also been applied in the derivation of CMs in vitro to increase the efficiency and maturation of cardiac differentiation. Collectively, a lot has been learned from the past failures of simply injecting intact stem cells or their derivatives in vivo for treating MI, and bioengineering stem cells with biomaterials may be a valuable strategy for advancing stem cell therapy towards its widespread application for MI treatment in the clinic.

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“…7. Working quickly, add the necessary volume of each Sendai reprogramming virus to the equilibrated medium 2 . This volume is determined by three variables -the number of cells, the titer of each virus, and the desired multiplicity of infection for each virus.…”

Section: Specialized Culture Techniquesmentioning

confidence: 99%

“…13. When colonies appear, stain the culture using sterile antibodies to confirm that the colonies are indeed pluripotent, as described previously 2 . As with feeding cells, ensure that the PSC media containing the antibodies has been equilibrated to the gases in the cell production facility.…”

Section: Specialized Culture Techniquesmentioning

confidence: 99%

Culturing Human Pluripotent and Neural Stem Cells in an Enclosed Cell Culture System for Basic and Preclinical Research

Stover

Herculian

Bañuelos

et al. 2016

JoVE

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This paper describes how to use a custom manufactured, commercially available enclosed cell culture system for basic and preclinical research. Biosafety cabinets (BSCs) and incubators have long been the standard for culturing and expanding cell lines for basic and preclinical research. However, as the focus of many stem cell laboratories shifts from basic research to clinical translation, additional requirements are needed of the cell culturing system. All processes must be well documented and have exceptional requirements for sterility and reproducibility. In traditional incubators, gas concentrations and temperatures widely fluctuate anytime the cells are removed for feeding, passaging, or other manipulations. Such interruptions contribute to an environment that is not the standard for cGMP and GLP guidelines. These interruptions must be minimized especially when cells are utilized for therapeutic purposes. The motivation to move from the standard BSC and incubator system to a closed system is that such interruptions can be made negligible. Closed systems provide a work space to feed and manipulate cell cultures and maintain them in a controlled environment where temperature and gas concentrations are consistent. This way, pluripotent and multipotent stem cells can be maintained at optimum health from the moment of their derivation all the way to their eventual use in therapy.

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Derivation of Induced Pluripotent Stem Cells by Lentiviral Transduction

Cited by 18 publications

References 7 publications

The Autism Spectrum Disorders Stem Cell Resource at Children's Hospital of Orange County: Implications for Disease Modeling and Drug Discovery

The Autism Spectrum Disorders Stem Cell Resource at Children's Hospital of Orange County: Implications for Disease Modeling and Drug Discovery

Stem Cell Therapy of Myocardial Infarction: A Promising Opportunity in Bioengineering

Culturing Human Pluripotent and Neural Stem Cells in an Enclosed Cell Culture System for Basic and Preclinical Research

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